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Gastric cancer (GC) constitutes substantial cancer mortality worldwide. Several cancer types aberrantly express bone marrow stromal cell antigen 2 (BST2), yet its functional and underlying mechanisms in GC progression remain unknown. In our study, RNA sequencing data revealed that BST2 was transcriptionally activated by homeobox D9 (HOXD9). BST2 was significantly upregulated in GC tissues and promoted epithelial-mesenchymal transition and metastasis of GC. BST2 knockdown reversed HOXD9's oncogenic effect on GC metastasis. Moreover, BST2 messenger RNA stability could be enhanced by poly(A) binding protein cytoplasmic 1 (PABPC1) through the interaction between BST2 3'-UTR and PABPC1 in GC cells. PABPC1 promoted GC metastasis, which BST2 silencing attenuated in vitro and in vivo. In addition, positive correlations among HOXD9, BST2, and PABPC1 were established in clinical samples. Taken together, increased expression of BST2 induced by HOXD9 synergizing with PABPC1 promoted GC cell migration and invasion capacity.
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Neoplasias Gástricas , Humanos , Neoplasias Gástricas/genética , Proteínas de Ligação a RNA , Movimento Celular/genética , Transição Epitelial-Mesenquimal/genética , RNA , Proliferação de Células , Linhagem Celular Tumoral , Regulação Neoplásica da Expressão Gênica , Metástase Neoplásica , Proteínas de Neoplasias , Proteínas de Homeodomínio/genética , Antígeno 2 do Estroma da Médula ÓsseaRESUMO
Age-related macular degeneration (AMD) is a leading blinding disease worldwide, and macular neovascularization (MNV) is a common complication encountered in the advanced stages of AMD. While the underlying causes of MNV remain elusive, aberrant multiplication of choroidal endothelial cells (CECs) and increased vascular endothelial growth factor (VEGF) are thought to play significant roles in the occurrence and development of MNV. Allograft inflammatory factor-1(AIF-1) is a crucial regulatory factor of vascular tubular structure formation and growth, involving the proliferation and migration of vascular endothelial cells and various tumor cells. This study aimed to understand how AIF-1 effects the proliferation of CECs and the subsequent progression of MNV. To study this, a mouse MNV model was established through laser injury, and the AIF-1 expression levels were then measured using western blot and immunohistochemistry. AIF-1 siRNA was intravitreally injected to silence AIF-1 gene expression. Western blot and choroidal flat mount were performed to measure the progression of MNV and proliferation of the CECs. These results showed that the protein expression of AIF-1 was significantly elevated in the laser-induced mouse MNV model, and the expression trend was consistent with VEGF. The protein level of AIF-1 was significantly decreased after the intravitreal injection of AIF-1 siRNA, the damage range of laser lesions was significantly reduced, and the proliferation of endothelial cells was inhibited. Knockdown of the AIF-1 gene significantly inhibited the expression of mitogen-activated protein kinase p44/42 in MNV lesions. In summary, this research demonstrates that AIF-1 promoted MNV progression by promoting the proliferation of CECs and that silencing AIF-1 significantly ameliorates MNV progression in mouse models, which may act through the p44/42 MAPK signaling pathway. AIF-1 could be a new potential molecular target for MNV.
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Neovascularização de Coroide , Degeneração Macular , Camundongos , Animais , Fator A de Crescimento do Endotélio Vascular/metabolismo , Células Endoteliais/metabolismo , Neovascularização de Coroide/metabolismo , Transdução de Sinais/fisiologia , RNA Interferente Pequeno/genética , Degeneração Macular/metabolismo , Proliferação de Células , LasersRESUMO
BACKGROUND: Lung cancer surgery is associated with a high incidence of postoperative pulmonary complications (PPCs). Preliminary evidence suggests that ERAS processes can reduce overall incidence of PPCs as short- and long-term recovery improved by supporting units to adopt evidence-based care. However, the evidence is inconclusive due to insufficient high-level studies in this research field. No well-designed, adequately powered, randomized controlled trials (RCTs) have investigated the effects of pulmonary rehabilitation based ERAS program (PREP) on post-operative pulmonary complications, pulmonary function, and health related quality of life following lung cancer surgery. METHODS: The PREP trial is a pragmatic, investigator-initiated, multi-center, randomized controlled, parallel group, clinical trial. Five hundred patients scheduled for minimally invasive pulmonary resection at six hospitals in China will be randomized with concealed allocation to receive either i) a pre-operative assessment and an information booklet or ii) a pre-operative assessment, an information booklet, plus an additional education, a 30-min pulmonary rehabilitation training session and the post-operative pulmonary rehabilitation program. The primary outcome is incidence of PPCs defined with the Melbourne Group Scale diagnostic scoring tool. Secondary outcomes include incidence of cardiopulmonary and other complications, pulmonary function, cardiopulmonary endurance, muscle strength, activity level, health-related quality of life (HRQoL), pre- and post-operative hospital length of stay (LOS), and total hospital LOS. DISCUSSION: The PREP trial is designed to verify the hypothesis that pulmonary rehabilitation based ERAS program reduces incidence of PPCs and improves pulmonary function and HRQoL in patients following lung cancer surgery. This trial will furthermore contribute significantly to the limited knowledge about the pulmonary rehabilitation based ERAS program following lung cancer surgery, and may thereby form the basis of future recommendations in the surgical community. TRIAL REGISTRATION: Chinese Clinical Trial Registry: ChiCTR1900024646, 21 July 2019.
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Neoplasias Pulmonares/reabilitação , Pulmão/fisiopatologia , Complicações Pós-Operatórias/epidemiologia , China , Humanos , Incidência , Neoplasias Pulmonares/cirurgia , Estudos Multicêntricos como Assunto , Ensaios Clínicos Pragmáticos como Assunto , Qualidade de Vida , Fatores de Risco , Resultado do TratamentoRESUMO
The persistent challenge of poor recovery characteristics of NO2 sensors operated at room temperature remains significant. However, the development of In2O3-based gas sensing materials provides a promising approach to accelerate response and recovery for sub-ppm of NO2 detection at room temperature. Herein, we propose a simple two-step method to synthesize a one-dimensional (1D) In2O3@ZnO heterostructure material with hollow microtubes, by coupling metal-organic frameworks (MOFs) (MIL-68 (In)) and zinc ions. Meanwhile, the In2O3@ZnO composite-based gas sensor exhibits superior sensitivity performance to NO2 under visible light activation. The response value to 5 ppm of NO2 at room temperature is as high as 1800, which is 35 times higher than that of the pure In2O3-based sensor. Additionally, the gas sensor based on the In2O3@ZnO heterostructure demonstrates a significantly reduced response/recovery time of 30 s/67 s compared to the sensor based on pure In2O3 (74 s/235 s). The outstanding gas sensing properties of the In2O3@ZnO heterostructure-based sensors can be attributed to the enhanced photogenerated charge separation efficiency resulting from the heterostructure effect, and the improved receptor function toward NO2, which can increase the reactive sites and gas adsorption capacity. In summary, this work proposes a low-cost and efficient method to synthesize a 1D heterostructure material with microtube structures, which can serve as a fundamental technique for developing high-performance room-temperature gas sensors.
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Índio , Luz , Dióxido de Nitrogênio , Temperatura , Óxido de Zinco , Óxido de Zinco/química , Dióxido de Nitrogênio/análise , Dióxido de Nitrogênio/química , Índio/química , Estruturas Metalorgânicas/química , Gases/análise , Gases/químicaRESUMO
Exhaled breath analysis has attracted considerable attention as a noninvasive and portable health diagnosis method due to numerous advantages, such as convenience, safety, simplicity, and avoidance of discomfort. Based on many studies, exhaled breath analysis is a promising medical detection technology capable of diagnosing different diseases by analyzing the concentration, type and other characteristics of specific gases. In the existing gas analysis technology, the electronic nose (eNose) analysis method has great advantages of high sensitivity, rapid response, real-time monitoring, ease of use and portability. Herein, this review is intended to provide an overview of the application of human exhaled breath components in disease diagnosis, existing breath testing technologies and the development and research status of electronic nose technology. In the electronic nose technology section, the three aspects of sensors, algorithms and existing systems are summarized in detail. Moreover, the related challenges and limitations involved in the abovementioned technologies are also discussed. Finally, the conclusion and perspective of eNose technology are presented.
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Dolichyl-diphosphooligosaccharide protein glycosyltransferase (DDOST) plays a pivotal role in the glycosylation of asparagine residues on nascent polypeptides. However, the biological role of DDOST in glioma remains unclear. The mRNA expression of DDOST in glioma was identified using TCGA, CGGA, GEO and Rembrandt datasets. Immunohistochemistry assay was conducted to examine the protein level of DDOST. Cox regression analysis, nomograms and calibration plots were used to evaluate the prognostic value of DDOST. The association between DDOST and immune cell infiltration was evaluated using CIBERSORT algorithm. Additionally, DNA methylation and ceRNA regulatory network of DDOST expression were investigated using the LinkedOmics and ENCORI databases. The authors found that DDOST was substantially expressed at the mRNA and protein levels. Functional enrichment analysis revealed close associations between DDOST and immune-related pathways, as well as immune cell infiltration. In addition, DDOST exhibited synergistic effects with tumour mutational burden (TMB) and other immune checkpoints. For expression regulation mechanisms, DDOST had low DNA methylation levels in high-grade gliomas and may be involved in multiple ceRNA networks in glioma. Thus, DDOST may serve as an unfavourable biomarker for gliomas. DNA methylation and ceRNA regulatory networks of DDOST expression were identified for the first time in this multi-omics study.
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Glioma , RNA , Humanos , Prognóstico , Multiômica , RNA Mensageiro , Glioma/diagnóstico , Glioma/genética , Glicosiltransferases/genéticaRESUMO
Retinal neovascularization (RNV) is a typical feature of ischemic retinal diseases that can lead to traction retinal detachment and even blindness in patients, in which the vascular endothelial cell growth factor (VEGF) plays a pivotal role. However, most anti-VEGF drugs currently used for treating RNV, such as ranibizumab, need frequent and repeated intravitreal injections due to their short intravitreal half-life, which increases the incidence of complications. Herein, a hydrogel intravitreal drug delivery system (DDS) is prepared by a dynamic Schiff base reaction between aminated hyaluronic acid and aldehyde-functionalized Pluronic 127 for sustained release of ranibizumab. The prepared hydrogel system named HP@Ran exhibits excellent injectability, self-healing ability, structural stability, cytocompatibility, and blood compatibility. According to an in vitro drug release study, the hydrogel system continuously releases the model drug bovine serum albumin for more than 56 days. Importantly, in an in vivo rabbit persistent RNV model, the HP@Ran hydrogel system continuously releases pharmacologically active ranibizumab for more than 7 weeks and also exhibits superior anti-angiogenic efficacy over ranibizumab treatment by decreasing vascular leakage and neovascularization at 12 weeks. Thus, the developed HP@Ran hydrogel system possesses great potential for intravitreal DDS for the treatment of RNV.
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Ranibizumab , Neovascularização Retiniana , Animais , Coelhos , Ranibizumab/farmacologia , Ranibizumab/uso terapêutico , Inibidores da Angiogênese/farmacologia , Inibidores da Angiogênese/uso terapêutico , Neovascularização Retiniana/tratamento farmacológico , Hidrogéis/química , Preparações de Ação Retardada/química , Biomimética , Fatores de Crescimento do Endotélio VascularRESUMO
PURPOSES: To evaluate the effects of low frequency whole-body vibration (WBV) on degeneration of articular cartilage and subchondral bone in mice with destabilization of the medial meniscus (DMM)induced osteoarthritis(OA) and mice with normal knee. METHODS: Ten-week-old C57BL/6J male mice received DMM on right knees, while the left knees performed sham operation. There were six groups: DMM, SHAM DMM, DMM+WBV,SHAM DMM+WBV, DMM+ NON-WBV and SHAM DMM+NON-WBV. After four weeks, the knees were harvested from the DMM and SHAM DMM group. The remaining groups were treated with WBV (10 Hz) or NON-WBV. Four weeks later, the knees were harvested. Genes, containing Aggrecan(Acan) and Collagenâ ¡(Col2a1), Matrix Metalloproteinases 3 and 13(MMP3,13), TNFα and IL6, were measured and staining was also performed. OA was graded with OARSI scores, and tibial plateaubone volume to tissue volume ratio(BV/TV), bone surface area to bone volume ratio (BS/BV), trabecular number(Tb.N) and trabecular thickness separation(TS) between groups were analyzed. RESULTS: Increased OARSI scores and cartilage degradation were observed after WBV. BV/TV, Tb.N and TS were not significant between the groups. Significant reductions were observed in MMP3, MMP13, Col2a1, Acan, TNFα and IL6 in the DMM+WBV compared to SHAM DMM+WBV group. BV/TV, BS/BV, Tb.N, TS and OARSI scores were not significantly changed in the left knees. IL6 expression in the SHAM DMM+WBV group was significantly increased compared with the SHAM DMM+ NON-WBV group, while Col2a1, Acan and MMP13 expression decreased. CONCLUSION: WBV accelerated cartilage degeneration and caused slight changes in subchondral bone in a DMM-induced OA model. WBV had no morphologic effect on normal joints.
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Cartilagem Articular , Osteoartrite do Joelho , Masculino , Animais , Camundongos , Camundongos Endogâmicos C57BL , Osteoartrite do Joelho/terapia , Fator de Necrose Tumoral alfa , Metaloproteinase 13 da Matriz/genética , Metaloproteinase 3 da Matriz , Interleucina-6/genética , Vibração/uso terapêuticoRESUMO
Transcription factors (TFs) and long noncoding RNAs (lncRNAs) contribute to gastric cancer (GC). However, the roles of TFs and lncRNAs in the invasion and metastasis of GC remain largely unknown. Here, we observed that the transcription factor VAX2 is significantly upregulated in GC cells and tissues and acts as an oncogene. Moreover, high VAX2 expression is associated with the advancement of tumors in GC. In terms of functionality, the enforced expression of VAX2 promotes the proliferation and metastasis of GC cells. Mechanistically, VAX2 specifically interacts with the LINC01189 promoter and represses LINC01189 transcription. Furthermore, LINC01189 exhibits significant downregulation in GC and functions as a suppressor gene. Functionally, it inhibits migratory and invasive abilities in GC cells. In the context of GC metastasis, VAX2 plays a role in modulating it by trans-repressing the expression of LINC01189. Additionally, LINC01189 binds to hnRNPF to enhance hnRNPF degradation through ubiquitination. The cooperation between LINC01189 and hnRNPF regulates GC cell invasion and migration. In addition, both VAX2 and hnRNPF are highly expressed, while LINC01189 is expressed in at low levels in GC tissues compared to normal gastric tissues. Our study suggests that VAX2 expression facilitates, while LINC01189 expression suppresses, metastasis and that the VAX2-LINC01189-hnRNPF axis plays a contributory role in GC development.
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PURPOSE: To explore the therapeutic efficacy of neuromuscular electrical stimulation (NMES) combined with swallowing rehabilitation training on the healing effect and quality of life of stroke patients with dysphagia. METHODS: The clinical data of 63 stroke patients admitted to the First Affiliated Hospital of Zhengzhou University from October 2019 to September 2020 were retrospectively analyzed. The included patients were divided into two groups according to different treatment plans: an observation group (n=33) treated with NMES combined with swallowing rehabilitation training, and a control group (n=30) treated by swallowing rehabilitation training alone. Before and after 2 courses of treatment, the Water swallow test, Functional Oral Intake Scale (FOIS), and MD Anderson Dysphagia Inventory (MDADI) were used to assess the swallowing function of patients in the two groups, and the National Institutes of Health Stroke Scale (NIHSS) was used to evaluate patients' neurological deficit; the SA7550 surface electromyogram (EMG) analysis system was applied to collect surface EMG, and the F113-5 medical X-ray TV system was used to detect the mobility of the hyoid-throat complex; the negative emotions of patients were assessed using the Hamilton Rating Scale for Depression (HAMD) before and after treatment, and the quality of life was evaluated by the Swallowing Quality of Life (SWAL-QOL) questionnaire; and the occurrence of adverse reactions during treatment was recorded and compared between the two groups. RESULTS: There was no significant difference in swallowing function, duration of swallowing, maximum amplitude value, and hyoid-throat complex mobility between the two groups before treatment (P>0.05), nor were there any differences in the scores of FOIS, MDADI, NIHSS, HAMD, and SWAL-QOL before treatment (P>0.05). After treatment, however, the above indicators of both groups were significantly improved (P<0.05), and the improvements were more significant in the observation group compared with the control group (P<0.05). Moreover, the incidence of adverse reactions in both groups were relatively low without significant difference between groups (P>0.05). CONCLUSION: NMES combined with swallowing rehabilitation training is effective in the treatment of swallowing dysfunction following stroke. It can effectively improve patients' swallowing function and quality of life, and relieve their negative emotions, with a high safety profile, which is worthy of clinical promotion.
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As an approach to coronary artery ostial injury in type A aortic dissection and infective endocarditis, we describe a technique of coronary ostial repair using a ring-shaped bovine pericardial patch. The inner and outer rims of the patch were sutured to the involved coronary ostium (to close the ostial tear) and to the aortic wall (to cover the sinus), respectively. Four patients were successfully managed and computed tomographic coronary arteriogram at follow-up showed patent coronary ostia and arteries. The favourable preliminary results imply that this technique is a simple, safe and effective approach to coronary ostial repair in patients with type A aortic dissection or infective endocarditis.
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Dissecção Aórtica , Procedimentos Cirúrgicos Cardíacos , Endocardite , Humanos , Bovinos , Animais , Vasos Coronários/diagnóstico por imagem , Vasos Coronários/cirurgia , Angiografia Coronária , Dissecção Aórtica/diagnóstico por imagem , Dissecção Aórtica/cirurgiaRESUMO
Background: Synovial macrophages play important roles in the formation and progression of osteoarthritis (OA). This study aimed to explore the biological and clinical significance of macrophage-associated genes (MAGs) in OA. Methods: The OA synovial gene expression profiles GSE89408 and GSE82107 were obtained from the GEO database. Single-sample gene set enrichment analysis (ssGSEA) and GSEA were employed to decipher differences in immune infiltration and macrophage-associated biological pathways, respectively. Protein-protein interaction (PPI) network analysis and machine learning were utilized to establish a macrophage-associated gene diagnostic signature (MAGDS). RT-qPCR was performed to test the expression of key MAGs in murine models. Results: OA synovium presented high levels of immune infiltration and activation of macrophage-associated biological pathways. A total of 55 differentially expressed MAGs were identified. Using PPI analysis and machine learning, a MAGDS consisting of IL1B, C5AR1, FCGR2B, IL10, IL6, and TYROBP was established for OA diagnosis (AUC = 0.910) and molecular pathological evaluation. Patients with high MAGDS scores may possess higher levels of immune infiltration and expression of matrix metalloproteinases (MMPs), implying poor biological alterations. The diagnostic value of MAGDS was also validated in an external cohort (AUC = 0.886). The expression of key MAGs was validated in a murine model using RT-qPCR. Additionally, a competitive endogenous RNA network was constructed to reveal the potential posttranscriptional regulatory mechanisms. Conclusions: We developed and validated a MAGDS model with the ability to accurately diagnose and characterize biological alterations in OA. The six key MAGs may also be latent targets for immunoregulatory therapy.
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Redes Reguladoras de Genes , Osteoartrite , Animais , Perfilação da Expressão Gênica , Humanos , Macrófagos/metabolismo , Camundongos , Osteoartrite/diagnóstico , Osteoartrite/genética , Osteoartrite/metabolismo , Membrana Sinovial/patologiaRESUMO
Complex oxide heterointerfaces provide a platform to manipulate spin-orbit coupling under the broken inversion symmetry. Moreover, their weak antilocalization (WAL) effect displays quantum coherent behavior due to the strong spin-orbit coupling. Herein, we break through the limitation of lattice mismatch at ReAlO3/STO (Re = La, Pr, Nd, Sm, and Gd) heterointerfaces and obtain their two-dimensional electric gas (2DEG) by spin coating. The effect of different Re elements in the resulting quantum corrections on the conductivity is investigated. It is observed that the conductivity of heterointerfaces is reduced with larger atomic numbers due to the ionization potential of Re elements. Moreover, magnetoresistance (MR) measurements in a perpendicular or a parallel field distinctly uncover strong Rashba spin-orbit coupling (SOC) in ReAO/STO samples besides SAO/STO (Re = Sm) and GAO/STO (Re = Gd), and the effective fields of the SOC (Hso) gradually increase from LAO/STO (Re = La, Hso = 0.82 T) to NAO/STO (Re = Nd, Hso = 1.37 T) at 2 K. The competition between SOC scattering and inelastic scattering is revealed through a temperature-dependence study of MR, and the WAL-weak localization transition is at about 6 K. Furthermore, unambiguous results of the Kondo effect, nonlinear Hall, hysteresis loop, and Rashba SOC suggest the coexistence of WAL at the PAO/STO (Re = Pr) heterointerface with exchange coupling between the localized magnetic moment and the itinerant electron. These results pave a unique route for the exploration of spin-polarized 2DEGs at oxide heterointerfaces.
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Although the amorphous two-dimensional electron gas (a-2DEG) of oxides provides new opportunities to explore nanoelectronic as well as quantum devices, the intrinsic effect of rare earth (Re = La, Pr, Nd, Sm, Gd, and Tm) elements at ReAlO3/SrTiO3 heterointerfaces is still largely unknown and needs to be addressed systematically. Herein, we first propose that the ionization potential of Re elements is a critical factor for the 2DEG fabricated by chemical spin coating. Furthermore, the photoresponsive properties of heterointerfaces are investigated comprehensively with the ionization potential ranging from 35.79 to 41.69 eV. The results show that the sheet resistances significantly increase with increasing the ionization potential, and a resistance upturn phenomenon is observed at TmAlO3/SrTiO3 heterointerfaces, which can be attributed to the weak localization effect theoretically. The most important observation is the dramatic transition from negative (-178.3%, Re = La) to positive (+89.9%, Re = Gd) photoresponse at ReAlO3/SrTiO3 heterointerfaces under the irradiation of 405 nm light at 50 K. More remarkably, a unique recovery behavior of transient-persistent photoconductivity coexistence at low temperatures is discovered at the TmAlO3/SrTiO3 heterointerface. This work reveals an effective approach to tune the transport and photoresponsive properties by changing Re elements and paves the way for the application of all-oxide devices.
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Raman spectroscopy yields a fingerprint spectrum and is of great importance in medical and biological sciences as it is non-destructive, non-invasive, and available in the aqueous environment. In this study, Raman spectroscopy and Raman mapping were used to explore the dynamic biochemical processes in screened bacteria under ceftazidime stress. The Raman spectral difference between bacteria with and without antibiotic stress was analyzed by principal component analysis and characteristic peaks were obtained. The results showed that amino acids changed first and lipids were reduced when bacteria were exposed to ceftazidime stress. Furthermore, in Raman mapping, when bacteria were subjected to antibiotic stress, the peak at 1002 cm-1 (phenylalanine) increased, while the peak at 1172 cm-1 (lipids) weakened. This indicates that when bacteria were stimulated by antibiotics, the intracellular lipids decreased and the content of specific amino acids increased. The reduction of intracellular lipids may suggest a change of membrane permeability. The increase of specific amino acids suggests that bacteria resist external stimuli of antibiotics by regulating the activities of related enzymes. This study explored the processes of the action between bacteria and antibiotics by Raman spectroscopy, and provides a foundation for the further study of the dynamics of microbial biochemical processes in the future.
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Magnetic two-dimensional electron gases at the oxide interfaces are always one of the key issues in spintronics, giving rise to intriguing magnetotransport properties. However, reports about magnetic two-dimensional electron gases remain elusive. Here, we obtain the magnetic order of LaAlO3/SrTiO3 systems by introducing magnetic dopants at the La site. The transport properties with a characteristic of metallic behavior at the interfaces are investigated. More significantly, magnetic-doped samples exhibit obvious magnetic hysteresis loops and the mobility is enhanced. Meanwhile, the photoresponsive experiments are realized by irradiating all samples with a 360 nm light. Compared to magnetism, the effects of dopants on photoresponsive and relaxation properties are negligible because the behavior originates from SrTiO3 substrates. This work paves a way for revealing and better controlling the magnetic properties of oxide heterointerfaces.
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OBJECTIVE: to study the effect of modified acellularization process on porcine endogenous retrovirus (PERV) in porcine aorta valves (PAVs). METHODS: Twenty aortic valves of pig were put into 0.1% trypsin solution, hypotonic and hypertonic TritonX-100, DNAse solution, RNAse solution, and Hanks solution in succession so as to remove the cells. The specimens of PAV were to undergo gross observation and microscopy before and after the acellularization procedure. Fracture test was made. Primers specific for the conservative gag gene of PERV were designed PCR and RT-PCR were used to detect the expression of gag. In addition, 20 samples of native PAV were collected. Peripheral mononuclear cells (PBMCs). Were isolated from 20 samples of porcine peripheral blood. Ten dogs underwent acellularized PAV replacement; 3 months later, samples of the dogs' peripheral blood were collected. Porcine kidney cells of the line PK15 were used as positive controls. RESULTS: Microscopy showed that all the cells were removed from the acellularized PAVs. Histological analysis showed that the major structural components were maintained. There was no significant difference in fracture strength between the native and acellularized PAVs (P > 0.05). PCR and RT-PCR showed a PERV 219 bp DNA fragment, 90%-95% homologous with the published PERV gene, in the genomic DNA of all native PAVs, pig PBMCs, and PK15 cells, but not in the acellularized PAVs and dog PBMCs. CONCLUSION: PERV exists in all native PAVs. The modified acellularization process succeeds in removing all the cell component and PERV in the PAVs, thus preventing cross-species transmission of PERV.
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Valva Aórtica/transplante , Valva Aórtica/virologia , Técnicas Citológicas/métodos , Retrovirus Endógenos , Animais , Linhagem Celular , Cães , Suínos , Engenharia Tecidual/métodos , Transplante HeterólogoRESUMO
OBJECTIVE: To explore the way of stably inducing canine bone marrow mesenchymal stem cells (BMSCs) to differentiate into fibroblasts and myofibroblasts in vitro, and provide seed cells for fabricating tissue engineering heart valves (TEHV). METHODS: Adult canine BMSCs were separated by a gradient centrifugation on Percoll (density 1.073 g/ml), then the cells were incubated in low-glucose Dulbecco Eagle's minimum essential medium (LG-DMEM) with 10% bovine calf serum. Cell phenotype were identified by immunohistochemistry staining. The second and third generation of BMSCs were committedly induced by conditioning culture medium, which were detected by immunohistochemistry staining. The induced-BMSCs were freezed, preserved and resuscitated after 7 d to observe the cell growth, proliferation and function. RESULTS: BMSCs deriving from the bone marrow mononuclear cells separated by a Percoll gradient were positive expression of alpha-smooth muscle antibody, vimentin and negative expression of CD34, laminin. About (50 +/- 3)% induced-BMSCs were positive expression of laminin. Approximately (85 +/- 3)% freezed induced-BMSCs could be resuscitated. And the growth, proliferation and function were well. CONCLUSION: BMSCs could be committedly induced to differentiate into fibroblasts and myofibroblasts in vitro. It is suitable to be the seed cells.